Target: TbPIP5K1 (phosphatidylinositol 4-phosphate 5-kinase) NCBI Gene # or RefSeq#:NCBI 185431 Protein ID (NP or XP #) or Wolbachia#:XP_844312.1. Organism (including strain):Trypanosoma brucei (strain 427) Etiologic Risk Group:BioSafety Level 2 Disease Information: Trypanosoma brucei is a parasitic protozoan that causes Human African Trypanosomiasis (HAT), commonly known as African sleeping sickness. Trypanosomiasis is primarily transmitted by the tsetse fly, and is thus endemic to sub-Saharan Africa. Trypanosomiasis symptoms include fever, headache, and adenopathy in the first stage, during which T. brucei invades the circulatory and lymphatic systems . This stage lasts approximately one to two weeks. If trypanosomiasis is left untreated in the first stage, the disease can be fatal in the second stage. During this stage, T. brucei invades the central nervous system and causes various neuropsychiatric symptoms, including tremors and eventually coma. The entire disease progression takes only approximately three weeks to manifest, ending in fatality.
Is it a monomer or multimer as biological unit? (make prediction at http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): Complex of proteins?: Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
PDB # or closest PDB entry if using homology model: 2YBX Current Inhibitors:unknown Expression Information:Yes, TbPIP5K1 has been successfully expressed in E. coli. Purification Method:Nickel resin tubes Image of protein:
Length: 481 amino acids Molecular Weight:54.379 kDa Molar Extinction coefficient of your protein at 280 nm wavelength: TMpred graph Image: CDS Gene Sequence: *GC% Content for gene: *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): *GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now) Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences): **
NCBI Gene # or RefSeq#: NCBI 185431
Protein ID (NP or XP #) or Wolbachia#: XP_844312.1.
Organism (including strain): Trypanosoma brucei (strain 427)
Etiologic Risk Group: BioSafety Level 2
Disease Information:
Trypanosoma brucei is a parasitic protozoan that causes Human African Trypanosomiasis (HAT), commonly known as African sleeping sickness. Trypanosomiasis is primarily transmitted by the tsetse fly, and is thus endemic to sub-Saharan Africa. Trypanosomiasis symptoms include fever, headache, and adenopathy in the first stage, during which T. brucei invades the circulatory and lymphatic systems . This stage lasts approximately one to two weeks. If trypanosomiasis is left untreated in the first stage, the disease can be fatal in the second stage. During this stage, T. brucei invades the central nervous system and causes various neuropsychiatric symptoms, including tremors and eventually coma. The entire disease progression takes only approximately three weeks to manifest, ending in fatality.
Link to TDR Targets page:
http://www.tdrtargets.org/targets/view?gene_id=16422
Link to Gene Database page:
http://tritrypdb.org/tritrypdb/showRecord.do?name=GeneRecordClasses.GeneRecordClass&source_id=Tb927.4.1620&project_id=TriTrypDB
Essentiality of this protein: Essential in T. brucei
http://www.sciencedirect.com/science/article/pii/S2451945616300939
Is it a monomer or multimer as biological unit? (make prediction at http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html):
Complex of proteins?:
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
EC#: 2.7.1.68
Link to BRENDA EC# page:
http://www.brenda-enzymes.org/enzyme.php?ecno=2.7.1.68
Figure 1: Reaction mechanism catalyzed by TbPIP5K1
Enzyme Assay information: Adenosine 5'-triphosphate Bioluminescence Assay Kit
PDB # or closest PDB entry if using homology model: 2YBX
Current Inhibitors: unknown
Expression Information: Yes, TbPIP5K1 has been successfully expressed in E. coli.
Purification Method: Nickel resin tubes
Image of protein:
Amino Acid Sequence:
Length: 481 amino acids
Molecular Weight: 54.379 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength:
TMpred graph Image:
CDS Gene Sequence:
*GC% Content for gene:
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
**